An Introduction to Metabolism Chapter 8
METABOLISM I. Introduction All of an organism s chemical reactions Thousands of reactions in a cell Example: digest starch use sugar for energy and to build new molecules
II. Metabolic Pathways starting molecule chemical reactions product(s) Each step catalyzed by specific enzyme Enzyme 1 Enzyme 2 Enzyme 3 A B C D Reaction 1 Reaction 2 Reaction 3 Starting Product molecule
Catabolic pathways - break down complex molecules into simpler compounds release energy for cells to use Ex. Glucose metabolism yeilds ATP
Anabolic pathways use energy to build complex molecules from simpler ones Ex. Protein synthesis from amino acids
III. Energy Energy capacity of a system to do work can be converted from one form to another 1. Kinetic energy = energy of motion Due to vibration of atoms /molecules
Kinetic energy examples: LIGHT Energy -electromagnetic energy includes visible light, x-rays, gamma rays and radio waves. MOTION energy in the movement of objects. The faster they move, the more energy. Wind is motion energy. When a car comes to a total stop, releases all motion energy in uncontrolled instant. SOUND -movement of energy through substances in longitudinal waves. Sound produced when force causes object or substance to vibrate energy is transferred through the substance in a wave. Far less than other forms of energy. HEAT average total kinetic energy of molecules
2. Potential energy = energy stored within system Ex. rubberband Chemical energy = PE available for release in chemical reaction Ex. Glucose has high CE
IV. The Laws of Energy Transformation Thermodynamics study of energy transformation In open system energy and matter can be transferred between system and surroundings Why are organisms/cells open systems?
First Law of Thermodynamics The energy of universe is constant: Energy can be transferred and transformed, but it cannot be created or destroyed. This is the principle of conservation of energy
Second Law of Thermodynamics Every energy transfer or transformation increases entropy (disorder) of the universe Living systems (open) increase entropy in environment During E transfer some E is released as heat (unusable)
Kinetic, potential energy Campbell Chemical energy Heat CO 2 + H 2 O (a) First law of thermodynamics (b) Second law of thermodynamics First Law: chemical E in food converted to kinetic E Second Law: disorder is entered into environment as heat, CO 2
V. Biological Order Cells create ordered structures from less ordered materials Ex. proteins built from amino acids Amino acids in cell -----------------------------------------------Growth hormone
Organisms replace ordered forms of matter and energy with less ordered break down molecules, releases heat Energy flows into ecosystem as and leaves as
VI. Free-energy (G) G = amt. of energy available to do work in biochemical reaction Δ G = change in free energy unstable systems move towards stable (equilibrium) System at equilibrium does no work
Equilibrium and Metabolism Reactions in closed system eventually reach equilibrium and then do no work G < 0 G = 0 (a) An isolated hydroelectric system It s a positive free energy change for the surroundings, negative for the system
Equilibrium and Metabolism Cells are open systems constant flow of materials not in equilibrium G < 0 (b) An open hydroelectric system
Equilibrium and Metabolism catabolic pathway releases free energy in series of reactions G < 0 G < 0 G < 0 (c) A multistep open hydroelectric system
Spontaneous processes 1. occur without (additional) energy input explosion, rusting (slow!) 2. release energy, usually heat 3. Gives up energy -Δ G 4. Exergonic The Gibbs free energy equation does depend on T, increasing T may increase reaction rate
Free energy -Δ G = exergonic reaction releases energy Spontaneous downhill Products have less G than reactants Exergonic reaction Reactants Energy Progress of the reaction (a)exergonic reaction: energy released Products Amount of energy released ( G < 0)
Cellular respiration Exergonic reaction C 6 H 12 O 6 + 6 O 2 6 CO 2 + 6 H 2 O ΔG = -686kcal/mol available for work ATP + H 2 O ADP + P i ΔG = -7.3 kcal/mol (hydrolysis)
+Δ G = endergonic rxn Absorbs free energy from surroundings Non-spontaneous
Many exergonic reaction are also exothermic (release heat) The bombardier beetle has two sets of chemicals stored separately in paired glands. The larger of the pair contains hydroquinones and H2O2. The the smaller gland contains catalases and peroxidases.
Photosynthesis Endergonic 6CO 2 + 6H 2 O (+ light energy) C 6 H 12 O 6 + 6O 2 686kcal/mol
Free energy Products Reactants Energy Amount of energy required ( G > 0) Progress of the reaction (b) Endergonic reaction: energy required
VI. ATP powers cellular work ATP High stored chemical potential energy drives cell activities
ATP ATP =adenosine triphosphate Energy released when terminal phosphate bond broken by hydrolysis
P P P Adenosine triphosphate (ATP) H 2 O P + P P + i Energy ATP --- ADP + P Δ G = -7.3kcal/mol
Phosphorylation Transfer P from ATP to other molecule to phosphorylate it. The phosphorylated molecule is less stable Phosphorylation opens an aquaporin (spinach) http://www.sciencedaily.com/releases/2005/12/051222085140.htm
The Nobel Prize in Physiology or Medicine 1992 Edmond H. Fischer and Edwin G. Krebs for their discoveries concerning "reversible protein phosphorylation as a biological regulatory mechanism".
Coupling = Use exergonic reactions to drive endergonic rxns (overall exergonic)
Regeneration of ATP Add phosphate group to ADP Muscle cell uses 10 million ATP per second ADP + P ATP is this + or - Δ G? Δ G = 7.3kcal/mol
Coupled reactions ATP + H 2 O Energy from catabolism (exergonic, energy-releasing processes) ADP + P i Energy for cellular work (endergonic, energy-consuming processes)
VIII. ENZYMES
A. Enzymes speed up metabolic reactions catalyst =chemical agent that speeds up a reaction Unchanged by reaction enzyme = catalytic protein (organic) Ex. Hydrolysis of sucrose by sucrase
B. Enzymes lower activation energy barrier chemical reaction = bonds broken/formed free energy of activation = activation energy (E a ) = energy to start chemical reaction E a contorts molecule makes bonds unstable No change in Δ G (doesn t affect reactants or product energy)
Course of reaction without enzyme Reactants Course of reaction with enzyme E A without enzyme E A with enzyme is lower G is unaffected by enzyme Progress of the reaction Products
C. How Enzymes Lower the E A 37 o C reactants do not reach E A (transition state) Enzymes speed rate to transition state Reactants most unstable Does not affect Δ G of rxn. Why?
D. Substrate Specificity of enzymes S= Substrate = reactant ES = Enzyme-substrate complex P = Product(s) Sucrose + Sucrase E-S complex Fructose + Glucose +? substrate enzyme products McGraw Hill Enzymes
Active site region on enzyme where substrate binds Induced fit - enzyme shape changes to fit to substrate Substrate Active site (a) Enzyme (b) Enzyme-substrate complex
E. Catalysis in Active Site 1000 1,000,000 rxns/sec Induced fit by: Hydrogen bonds Ionic bonds Hydrophobic interactions Enzyme can catalyze forward or reverse rxn Always in direction of equilibrium Active site can lower E a Orient substrates Contort bonds Provide microenvironment Covalently bond to substrate
Substrates enter active site; enzyme changes shape such that its active site enfolds the substrates (induced fit). Substrates held in active site by weak interactions, such as hydrogen bonds and ionic bonds. Active site is available for two new substrate molecules. Substrates Enzyme Enzyme-substrate complex Active site can lower E A and speed up a reaction. Products are released. Products Substrates are converted to products.
F. Local Conditions activity affected by temperature, ph (each has optimal) chemicals
Rate of reaction Rate of reaction Optimal temperature for typical human enzyme Optimal temperature for enzyme of thermophilic (heat-tolerant) bacteria 0 20 40 60 80 100 Temperature (ºC) (a) Optimal temperature for two enzymes Optimal ph for pepsin (stomach enzyme) Optimal ph for trypsin (intestinal enzyme) 0 1 2 3 4 5 6 7 8 9 10 ph (b) Optimal ph for two enzymes
G. Cofactor assists enzyme Non-protein inorganic cofactor Metal ion (trace elements Zn 2+ (cofactor for carbonic anhydrase), Cu 2+, Mn 2+, K +, Na +.) organic cofactor is a coenzyme Vitamins (B, C, K) Bind to active site
H. Enzyme Inhibitors Competitive inhibitors bind to active site of enzyme compete with substrate at active site Alcohol dehydrogenase enzyme
Noncompetitive inhibitors bind to another part of enzyme change shape of active site May not be reversible toxins, poisons, pesticides, antibiotics
Substrate Active site Competitive inhibitor Enzyme (a) Normal binding (b) Competitive inhibition Noncompetitive inhibitor (c) Noncompetitive inhibition
9. Regulation of enzyme activity helps control metabolism metabolic pathways are tightly regulated
Allosteric Regulation (normal in cells) Allosteric enzyme with four subunits Active site (one of four) Regulatory molecule binds to enzyme but not at the active sites! (note: each subunit has its own active site) Regulatory site (one of four) Oscillation Active form Most enzymes have inactive and active forms shape changers! Nonfunctional active site Inactive form
Allosteric Regulation A. Allosteric inhibition molecule binds to stabilize inactive state blocks substrate Binding is reversible Nonfunctional active site Inactive form Inhibitor Stabilized inactive form
B. Allosteric cooperativity boosts enzyme activity Regulatory molecule binds to one site stabilizes favorable shape changes at other subunits Regulatory site (one of four) Active site (one of four) Active form Activator Stabilized active form
IX. Feedback Inhibition End product of metabolic pathway turns off pathway No wasting chemical resources by synthesizing more product than is needed
Initial substrate (threonine) Isoleucine used up by cell Isoleucine binds to allosteric site Feedback inhibition Active site available Active site of enzyme 1 no longer binds threonine; pathway is switched off. Intermediate A Enzyme 2 Intermediate B Enzyme 3 Intermediate C Enzyme 4 Intermediate D Enzyme 5 Threonine in active site Enzyme 1 (threonine deaminase) End product (isoleucine)